Division of Nephrology, Department of Internal Medicine III, J.W. Goethe-University, Frankfurt/M, Germany.
Stem Cells Dev. 2013 Jan 15;22(2):330-9. doi: 10.1089/scd.2012.0346. Epub 2012 Oct 4.
The characterization of adipose-derived stromal/stem cells (ASCs) remains difficult due to the lack of a definitive and unique cellular marker. Therefore, a combination of markers is necessary to identify the cells. No comprehensive analysis of the immunophenotype of expanded plastic adherent ASCs has been published. Therefore, the aim of this study was to characterize the general phenotype of cultured ASCs and to further analyze cellular subsets. ASCs were isolated from lipoaspirates from patients undergoing cosmetic liposuction and cultured in standard cell culture. A comprehensive phenotype characterization was done with the BD Lyoplate™ Human Cell Surface Marker Screening Panel containing 242 antibodies and isotype controls. Cultured ASCs not only showed the characteristic expression profile of mesenchymal stem cells (MSCs), but also revealed donor-specific variability in the expression of 49 other markers. We further detected markers with a scattering in the fluorescence intensity, indicating subpopulations with different expression profiles. Therefore, a multi-color flow cytometric analysis was done after staining the cells with direct-labeled antibodies against CD73, CD90, CD105, and either CD34, CD140b, CD200, CD201, or CD36 to verify the selected subpopulations of ASCs. We detected no CD34-CD36 double-positive population, but CD34(+)-CD36(-) and CD34(-)CD36(+) subpopulations, both of which are positive for the 3 main MSC markers, CD73, CD90, and CD105. All other detected subpopulations also co-expressed the 3 main MSC markers, and therefore fulfill the minimal phenotypic criteria for the definition of cultured MSCs. Our study demonstrates the first comprehensive phenotypic characterization of ASCs and clearly highlights donor-specific variability in ASC preparations.
脂肪来源的基质/干细胞 (ASC) 的特征仍然很困难，因为缺乏明确和独特的细胞标记物。因此，需要组合标记物来识别细胞。尚未发表关于扩增的塑料贴壁 ASC 的免疫表型的综合分析。因此，本研究的目的是表征培养的 ASC 的一般表型，并进一步分析细胞亚群。从接受美容抽脂的患者的脂肪抽吸物中分离 ASC，并在标准细胞培养中培养。使用包含 242 种抗体和同型对照的 BD Lyoplate™ 人类细胞表面标记物筛选试剂盒进行全面的表型特征描述。培养的 ASC 不仅表现出间充质干细胞 (MSC) 的特征表达谱，而且还显示出供体特异性的 49 种其他标记物表达的可变性。我们进一步检测了荧光强度散射的标记物，表明存在具有不同表达谱的亚群。因此，在用针对 CD73、CD90、CD105 的直接标记抗体对细胞进行染色后，进行了多色流式细胞术分析，以验证所选的 ASC 亚群。我们未检测到 CD34-CD36 双阳性群体，但检测到 CD34(+)-CD36(-)和 CD34(-)CD36(+)亚群，这两个亚群均对 3 种主要 MSC 标记物 CD73、CD90 和 CD105 呈阳性。所有其他检测到的亚群也共同表达了 3 种主要 MSC 标记物，因此符合培养 MSC 的定义的最小表型标准。我们的研究首次全面表征了 ASC，并清楚地突出了 ASC 制剂中的供体特异性可变性。
